Histopathological, morphological, and molecular characterization of fish-borne zoonotic parasite Eustrongylides Excisus infecting Northern pike (Esox lucius) in Iran.

Eustrongylides excisus is a fish-borne zoonotic parasite known to infect various fish species, including Northern pike (Esox Lucius). This nematode, belonging to the family Dioctophymatidae, has a complex life cycle involving multiple hosts. This study aimed to investigate the occurrence of Eustrongylides nematodes in Northern pike (E. Lucius) collected from Mijran Dam (Ramsar, Iran). Between June and October 2023, an investigation was conducted on Northern pike from Mijran Dam in Ramsar, Iran, following reports of reddish parasites in their muscle tissues. Sixty fish were examined at the University of Tehran, revealing live parasites in the muscles, which were then analyzed microscopically and preserved for a multidisciplinary study. The skeletal muscle tissues of 85% (51/60) of fish specimens were infected by grossly visible larvae which were microscopically identified as Eustrongylides spp. In histopathological examination, the lesion was composed of encapsulated parasitic granulomatous myositis. Microscopically, the cystic parasitic granulomas compressed the adjacent muscle fibers, leading to their atrophy and Zenker's necrosis. Moreover, epithelioid macrophages, giant cells and mononuclear inflammatory cells were present around the larvae and between the muscle fibers. Finally, a molecular analysis by examining the ITS gene region, revealed that they belong to the species E. excisus. Eustrongylidiasis in northern Iran necessitates further research into the biology, epidemiology, and control of Eustrongylides nematodes, focusing on various hosts. This study is the first to comprehensively characterize E. excisus in Northern pike in Ramsar, Iran, raising concerns about possible zoonotic transmission.

Lumpy jaw caused by Nocardia farcinica in a Holstein cow.

A 3-year-old Holstein cow was examined in an intensive system due to unilateral swelling in the mandible. A right mandibular mass was associated with painful mastication and Ptyalism. In palpation, the mass was raised, ulcerated, attached to the mandible bone and firm, approximately 17 × 12 × 10 cm3 in size. The lesion was sampled, and after routine bacteriology and histopathology procedures, the occurrence of lumpy jaw caused by Nocardia farcinica was confirmed. The bacterium was analysed using genome sequencing and new strain called Najm 114. Due to the risk of zoonosis of the isolated agent, the cow was euthanized. This is the first report of lumpy jaw caused by N. farcinica in a cow. This study showed that N. farcinica should be considered a possible etiological agent for lumpy jaw in cattle.

Molecular identification, phylogenetic analysis and histopathological study of pathogenic free-living amoebae isolated from discus fish (Symphysodon aequifasciatus) in Iran: 2020-2022.

Free-living amoebae (FLA) are capable of inhabiting diverse reservoirs independently, without relying on a host organism, hence their designation as "free-living". The majority of amoebae that infect freshwater or marine fish are amphizoic, or free-living forms that may colonize fish under particular circumstances. Symphysodon aequifasciatus, commonly referred to as the discus, is widely recognized as a popular ornamental fish species. The primary objective of the present study was to determine the presence of pathogenic free-living amoebae (FLA) in samples of discus fish. Fish exhibiting clinical signs, sourced from various fish farms, were transferred to the ornamental fish clinic. The skin, gills, and intestinal mucosa of the fish were collected and subjected to culturing on plates containing a 1% non-nutrient agar medium. The detection of FLA was conducted through morphological, histopathological and molecular methods. The construction of the phylogenetic tree for Acanthamoeba genotypes was achieved using the maximum likelihood approach. The molecular sequence analysis revealed that all cultures that tested positive for FLA were T4 genotype of Acanthamoeba and Acanthamoeba sp. The examination of gill samples using histopathological methods demonstrated the presence of lamellar epithelial hyperplasia, significant fusion of secondary lamellae, and infiltration of inflammatory cells. A multitude of cysts, varying in shape from circular to elliptical, were observed within the gills. The occurrence of interlamellar vesicles and amoeboid organisms could be observed within the epithelial tissue of the gills. In the current study, presence of the Acanthamoeba T4 genotype on the skin and gills of discus fish exhibiting signs of illness in freshwater ornamental fish farms was identified. This observation suggests the potential of a transmission of amoebic infection from ornamental fish to humans, thereby highlighting the need for further investigation into this infection among ornamental fish maintained as pets, as well as individuals who interact with them and their environment.

Acute coenurosis in lambs.

Six 100-day-old mixed-breed lambs were examined in a farm with a semi-intensive system due to neurologic signs. Cachexia, bilateral blindness, stupor, severe drowsiness and lethargy with left and right movements of the head and neck were recorded after awakening and stimulation. Lambs died 10 days after the onset of the clinical signs. The lambs were necropsied, and after routine parasitology, bacteriology and histopathology, the occurrence of acute coenurosis was confirmed due to finding multiple cystic structures in the brain tissue. All lambs of the herd were treated with albendazole (orally, 25 mg/kg, two doses with an interval of 14 days). All shepherd dogs were treated with popantel (orally, one tablet/10 kg, two doses with an interval of 14 days). The affected lambs died despite this treatment. No new case of the disease was observed after the initiation of control measures. The present study shows the importance of preventive measure against coenurosis in a semi-intensive sheep farming system that includes implementing consistent parasite control programme in dogs being in contact with sheep.

Clinico-histopathological and phylogenetic analysis of protozoan epibiont Epistylis wuhanensis associated with crustacean parasite Lernaea cyprinacea from ornamental fish in Iran.

Parasitic infestations are one of the most concerning problems limiting ornamental fish farming. In addition to the direct economic losses due to the major mortalities, parasites may significantly negatively impact the body shape, coloration, growth rate, and health condition of the fish. The results of the study highlight the importance of comprehensive parasitological analysis in the diagnosis and treatment of parasitic infections in ornamental fish farms. The presence of multiple parasites in the affected fish emphasizes the need for effective biosecurity measures, such as extending the quarantine period for newly imported fish, closely monitoring fish populations, and implementing isolation units to prevent the spread of infections. By implementing these preventative measures, ornamental fish farmers can reduce the risk of parasitic infections and ensure the health and well-being of their fish populations. This, in turn, can lead to increased profitability and sustainability for their business. Overall, the current study aimed to conduct a clinical, histopathological, and phylogenetic analysis of the epibiont ciliated protozoan Epistylis wuhanensis and the copepod crustacean Lernaea cyprinacea in a freshwater ornamental fish farm in Iran. Furthermore, it provides valuable insights into the prevalence and impact of parasitic infections in ornamental fish farms and underscores the need for continued research and the development of effective preventative measures to address this issue. A total of 60 symptomatic freshwater ornamental fish, including 30 guppy (Poecilia reticulata) and 30 sailfin molly (Poecilia latipinna), were packed in polyethylene bags filled with oxygenated pond water and transported to the Ornamental Fish Clinic, Faculty of Veterinary Medicine, University of Tehran, for parasitological analysis. Following the clinical examination, histopathological analysis was performed on 10% NBF (neutral buffered formalin)-fixed samples from affected tissues, including the skin, skeletal muscle, and liver, to identify any pathological changes associated with the parasitic infections. Furthermore, the DNA was extracted from the 99% ethanol-fixed samples using a commercial DNA extraction tissue kit (SinaPure DNA, Iran), and PCR was performed using Peri18S-F1 (5'-ACC TGG TTG ATC CTG CCA GT-3') and Peri18S-R1 (5'-TGC AGG TTC ACC TAC GGA AA-3') (first reaction), and Peri18S-F2 (5'-CCG CGG TAA TTC CAG CTC-3') and Peri18S-R2 (5'-GAT CCC CTA ACT TTC GTT CTT GA-3') (second round) primers for the identified parasites. Finally, the PCR products were sequenced using Sanger dideoxy sequencing methods, and the resulting sequences were compared to sequences in the BLAST search program to provide a comprehensive picture of the current parasite-based disorder. The crustacean L. cyprinacea and the epibiont sessilid E. wuhanensis were identified in the examined ornamental guppy (6/30) and sailfin molly (6/30), with an overall parasitic prevalence of 20.00% (12/60). Ciliates were found in all tissue lesions but not in fish without lesions. A great number of the ciliated protozoan E. wuhanensis were found attached to the integumentary area of L. cyprinacea. Microscopically, oval to round granulomatous lesions were observed in cutaneous and skeletal muscles. Lymphoplasmacytic dermatitis and myositis were also observed. The crustacean L. cyprinacea serves as a mechanical vector for E. wuhanensis infection and spreads the disease in ornamental fish farming operations. For the first time in Iran, we successfully presented diagnostic morphological and molecular data on sessilids isolated from L. cyprinacea. Based on the findings of the current study, such parasitic infections may cause significant economic losses following invasion of the integument area of fish, eventually leading to death, if treatment is neglected or inadequate. Furthermore, the findings of the analysis were used to develop effective diagnostic approaches for the affected fish, as well as recommendations for improved health conditions to prevent future outbreaks of parasitic infections. However, further research is needed to determine the precise mechanisms of crustacean attachment and host-crustacean-peritrich protozoan interactions. Furthermore, the direct and indirect effects of various environmental factors on the emergence and spread of the current disease should be considered.

The study of a hermaphroditic sheep caused by a mutation in the promoter of SRY gene.

In mammals, sex-determining region Y (SRY) gene plays vital role as a transcription factor to regulate the expression of the genes contributing to development of male genitals. Any mutation disrupting expression of SRY gene can cause disorders of sex development (DSDs). In this study, the examination of a hermaphroditic (female-like) Shal sheep which was referred for infertility is described. Initially, the reproductive system of the sheep was histologically and anatomically assessed. Karyotyping was used to determine the real gender of the animal. Sex hormones including progesterone, estradiol, and testosterone were measured by enzyme-linked immunosorbent assay (ELISA). Eventually, promoter part and SRY gene were sequenced and aligned to detect any potential mutation using NCBI data base. Although anatomical inspection led to identification of uterus, ovary, and enlarged clitoris as well as testes in the sheep, the karyotyping results interestingly revealed that the animal was genetically a male. Although the sheep had both male and female gonads, there were no overt signs of reproductive behavior and gamete production was not observed. Plasma steroid hormone levels were reported to be at basal levels. Additionally, a mutation was detected on the promoter of the SRY gene. In conclusion, the case implies that mutation on the promoter part of SRY gene could disrupt sexual development of the fetus culminating in DSDs in the sheep.

The effect of subacute co-exposure to carbon tetrachloride and diclofenac on the liver of male wistar rats.

Carbon tetrachloride (CCl4) is a potent liver toxin. Diclofenac (Dic), leading adverse effects on the liver, is used among the employees of the industries that use CCl4. The increased use of CCl4 and Dic in industrial workers has prompted us to investigate their synergistic effect on the liver using male Wistar rats as a model. Male Wistar rats were divided into seven groups (n = 6), and the exposure was by intraperitoneal injection for 14 days as follows. Group 1: Control, 2: Olive oil, 3: CCl4 (0.8 mL/kg/day (3 times per week)), 4: Normal saline, 5: Dic (1.5 mg/kg/day per day), 6: Olive oil with normal saline, 7: CCl4 (0.8 mL/kg/day (3 times per week)) and Dic (1.5 mg/kg/day daily). At the end of day 14, the heart blood was collected to measure the liver enzymes, alanine-aminotransferase (ALT), aspartate-aminotransferase (AST), blood alkaline phosphatase (ALP), albumin (ALB), direct bilirubin, and total bilirubin. A pathologist examined the liver tissue. Prism software was used to analyze data using ANOVA and Tukey statistical tests. ALT, AST, ALP, and Total Bilirubin enzymes were increased significantly in the co-administered CCl4 and Dic group, while the ALB levels decreased (p < 0.05). The histological findings reported liver necrosis, focal hemorrhage, adipose tissue change, and lymphocytic portal hepatitis. In conclusion, using Dic while exposed to CCl4 may exacerbate hepatotoxicity in rats. Therefore, it is suggested that more severe restrictions and safety regulations be placed on using CCl4 in the industry, and caution is advised to these industry workers to use Diclofenac.

Acute cysticercosis caused by Cysticercus tenuicollis in lambs.

Twelve 30- to 75-day-old mixed breed lambs were examined in an intensive system because of sudden recumbency and death. Clinical examination revealed sudden recumbency, visceral pain, and auscultation of respiratory crackles. Lambs died shortly (between 30 minutes and 3 hours) after the onset of clinical signs. The lambs were necropsied, and after routine parasitology, bacteriology and histopathology procedures, the occurrence of acute cysticercosis caused by Cysticercus tenuicollis was confirmed. The use of the suspect infested feed (newly purchased starter concentrate) was discontinued and other lambs of the flock were treated with praziquantel (15 mg/kg, single dose, orally). After these actions, no new cases were observed. The present study showed the importance of preventive measures against cysticercosis in intensive sheep farming systems which include proper storage of feed, preventing feed and environmental access by potential definitive hosts, and implementing consistent parasite control programs in dogs that are in contact with sheep.

Pericarditis caused by Proteus mirabilis in sheep.

Pericarditis means inflammation in the pericardial sac. Pericarditis is divided into three categories based on morphology, including fibrinous, purulent, and constrictive. In the present study, a 7-month-old male Ghezel breed sheep was examined for low weighting rate for three months. Tachypnea, tachycardia, heart friction sound, absence of fever and normal appetite were recorded in the clinical examination. In the patient's history, there was a history of perforated chest trauma behind the left scapula about three months ago. After the echocardiography examination and bacteriology procedures, purulent pericarditis caused by Proteus mirabilis was diagnosed. The bacterium was analysed using genome sequencing and new strain called Abhar114 was diagnosed. This is the first report of pericarditis caused by Proteus mirabilis in sheep.

Effect of L-serine on oxidative stress markers in the kidney of streptozotocin-induced diabetic mice.

Oxidative stress is critical in the occurrence and development of diabetes and its related complications. L-serine has recently been shown to reduce oxidative stress, the incidence of autoimmune diabetes and improve glucose homeostasis. The aim of this study was to investigate the effects of daily L-serine administration on blood glucose, renal function and oxidative stress markers in the kidney of streptozotocin-induced diabetic mice. Eighteen C57BL/6 male mice were randomly divided into three groups (n = 6 per group). Streptozotocin was used to induce diabetes and a group of diabetic mice was treated with 280 mg/day of L-serine dissolved in drinking water for 4 weeks. The level of blood glucose, biochemical markers of renal function (total protein, urea, creatinine and albumin) and oxidative stress markers (protein carbonyls, malondialdehyde, glutathione peroxidase, superoxide dismutase and catalase) were measured using spectrophotometry. The results indicated that L-serine significantly decreased the glucose level in diabetic mice (188.6 ± 22.69 mg/dL, P = 0.02). Moreover, treatment of diabetic mice with L-serine reduced protein carbonyls (3.249 ± 0.9165 nmol/mg protein, P < 0.05) and malondialdehyde levels (1.891 ± 0.7696 µM/mg protein, P = 0.051). However, L-serine showed no significant effects on renal function, and a slight reduction in histopathological changes was observed in mice receiving L-serine. This study revealed that L-serine effectively ameliorates oxidative stress in kidney tissue and reduces the blood glucose concentration in diabetic mice.

A rare case of ocular and testicular T-cell lymphoma in a hermaphrodite koi carp (Cyprinus carpio Linnaeus 1758): clinical, histopathological, and immunohistochemical study.

BACKGROUND: Lymphatic neoplasia may occur in various types, such as lymphoma, lymphosarcoma, lympholeukemia, and plasmacytoid leukemia. Lymphoma, defined as a malignant tumour of lymphoid tissue, has been found in a number of fish families including Esocidae and Salmonidae. However, the occurrence of lymphoma is rare in those belonging to the Cyprinidae. A final diagnosis of ocular and testicular T-cell lymphoma in the present study was based on the clinical signs, morphology, and texture of the tumour masses in the macroscopic and microscopic examinations. In addition, histopathological and immunohistochemical findings corresponded to T-cell lymphoma characteristics. CASE PRESENTATION: A 2-year-old hermaphrodite koi carp (Cyprinus carpio Linnaeus 1758) with a large ocular mass and severe exophthalmia in the right eye was referred to the Ornamental Fish Clinic in October 2020. Under anesthesia, enucleation was performed. 57 days after enucleation of the right eye, exophthalmia in the left eye was discovered. 221 days after surgery, the fish was discovered to be dead. At necropsy, a large soft tissue mass attached to the left testis was discovered. There were also small whitish nodules on the surface of the liver. Histopathology revealed a hypercellular ocular mass with scant connective tissue. The sections also revealed multifocal hemorrhages, round to ovoid neoplastic cells, mild-to-moderate anisokaryosis and anisocytosis, and mitotic figures. Basophilic neoplastic cells were found in blood vessels within the testicular mass, raising the possibility of systemic spread. The liver showed microscopic metastasis with morphologic similarities to the ocular and testicular tumors. The neoplastic cells infiltrating the left and right eyes as well as the testicular mass were immunohistochemically positive for CD3 but negative for CD20. The masses were diagnosed as T-cell lymphoma based on histopathological and immunohistochemical findings. CONCLUSIONS: This case report provides the first evidence of clinical, histopathological, morphological, and immunohistochemical findings of an ocular and testicular T-cell lymphoma in a hermaphrodite koi carp (Cyprinus carpio) in Iran.

The anti-inflammatory effects of minocycline on lipopolysaccharide-induced paw oedema in rats: a histopathological and molecular study.

Minocycline is a semi-synthetic antimicrobial agent with claimed anti-inflammatory properties reported from different experimental models. This study was aimed to evaluate the anti-inflammatory effects of minocycline, compared to the actions of two common anti-inflammatory agents, on lipopolysaccharide (LPS)-induced paw oedema through some clinical, histopathological, haematological and molecular analyses. Forty-eight rats were divided into eight groups (n = 6). In control group (Ctrl), each animal was injected with normal saline into its sub-plantar region of hind paw. In groups 2-7, hind paw oedema was induced by injection of LPS. One hour before injections, groups 1 (Ctrl) and 2 (LPS) were treated orally with distilled water, 3 and 4 with methylprednisolone (Pred) and meloxicam (Melo) and 5-7 with minocycline in doses of 50, 150 and 450 mg/kg (M50, M150 and M450, respectively). The 8th group (MC) was given minocycline (150 mg/kg) orally and normal saline was injected into sub-plantar region. Paw swelling and body temperature were assessed at 0, 2, 4, 6 and 24 h post-injections. At 24 h, samples of blood and liver, kidney, spleen and hind paw tissues were taken for haematological and histopathological examinations. Some samples of the paw were also obtained for molecular analysis of some inflammatory-related cytokines at mRNA level. Paw swelling and body temperature increased in all LPS-injected groups 2 h post-injection. In LPS group, they remained significantly increased up to 24 h; however, these parameters decreased to normal in Pred, Melo and all minocycline groups. The histological findings showed mild-to-moderate signs of inflammation in tissue samples of groups 2-6, but not in group M450. Additionally, gene expression of pro-inflammatory cytokines (IL-1ß and IL-6) increased significantly in LPS group compared to other groups. In conclusion, this study supports the role of minocycline as an anti-inflammatory agent with effects comparable to those of meloxicam and methylprednisolone.

First identification of Mycoplasma capricolum subspecies capripneumoniae in goats in Iran.

Mycoplasma capricolum subspecies capripneumoniae (Mccp) is the etiological agent of caprine contagious pleuropneumonia (CCPP) disease. The CCPP is one of the most severe diseases of goats. A herd of 2,000 goats located in the countryside of Tehran city, Iran, was examined for the study in August 2021. In history taking, observation, inspection and clinical examination, high case fatality rate (46.00%) due to respiratory distress and high morbidity of pleuropneumonia (15.00%) syndrome were recorded. Accordingly, ten carcasses of goats were dissected. The epidemiological pattern of the disease, clinical examination findings and the signs of necropsy of dead patients were suspected to CCPP. Four lung samples of necropsied goats were sent to the laboratory for PCR test and in all of them, Mccp was detected and CCPP was also confirmed. The disease was controlled by two measures: (a) the whole herd was first treated with antibiotics (florfenicol and tylosin) and (b) then the Pulmovac-In vaccine was then administered. This study is the first documented report of CCPP occurrence caused by Mccp in Iran and shows the importance of availability of effective vaccines to control and prevention of CCPP.

Histological observations on aural fibrosarcoma in a Holstein cow.

Fibrosarcomas occur as a mesenchymal tumour of malignant fibroblasts in a collagen background and are usually found in the female genital organs and rarely involve the skin. A 5-year-old female Holstein cow with a raised mass at the base of right ear was referred. On gross examination, the mass was approximately 13.00 × 10.00 × 7.00 cm in size. Finally, complete surgical removal was selected. The mass was encapsulated and the dermis was expanded by spindle-shaped to polygonal neoplastic cells. These cells were arranged in interwoven pattern. Mitotic figures were infrequent. Masson's trichrome demonstrated the positive and blue staining of collagen. Immunohistochemically, the sections were uniformly positive for Vimentin and negative for Desmin, SMA and GFAP. A well-differentiated fibrosarcoma was diagnosed based on histopathological features. Surgical excision is the treatment of choice for this neoplasm. In the present case, surgery was also performed successfully and no new growth of the mass was observed 4 months following the surgical procedures. To our knowledge, this is the first report of well-differentiated fibrosarcoma in a Holstein cow.

The immunomodulatory effect of minocycline on gene expression of inflammation related cytokines in lipopolysaccharide-treated human peripheral blood mononuclear cells.

To evaluate the immunomodulatory effect of minocycline, the present study was carried out on the gene expression of toll-like receptor type-4 (TLR4) and some pro-inflammatory (IL-1ß, IL-6) and anti-inflammatory cytokines (IL-10) associated with lipopolysaccharide (LPS) -induced inflammation in human peripheral blood mononuclear cells (PBMCs). The PBMCs were collected and then 5.4 × 106 PBMCs/mL were used in eight groups as follows: control group (only media), LPS group (only LPS), methylprednisolone (Pred) group (LPS plus Pred), meloxicam (Melo) group (LPS plus Melo), three minocycline groups [M1, M5 and M25] (LPS plus 1, 5, and 25 µg/mL minocycline, respectively) and minocycline control (MC) group (5 µg/mL minocycline). After incubation for 24 h, the PBMCs were subjected to quantitative PCR assays. Gene expression levels of TLR4 were not changed in any groups. The IL-1ß levels were increased in the LPS group but the increases were much more intense in the other groups except Pred group. Compared with control group, IL-6 levels increased significantly in Melo, M1 and M25 groups. Significant increases of IL-10 levels were also observed in Melo, M25 and MC groups. It can be concluded that minocycline had dual pro- and anti-inflammatory activities with potential clinical immunomodulatory effects.

Pyoderma caused by Proteus mirabilis in sheep.

Pyoderma is a purulent skin infection usually caused by bacteria and can be divided into primary and secondary categories based on histology. In the present study, an 18-month-old female mixed breed sheep was examined for pyoderma at the injection site of the enterotoxemia vaccine. After routine bacteriology and histopathology procedures, secondary pyoderma caused by Proteus mirabilis was diagnosed. The bacterium analysed using genome sequencing and new strain called AJJ 2021 was diagnosed. This is the first report of pyoderma caused by Proteus mirabilis in sheep.

Basal cell carcinoma in an albino pindani (Chindongo socolofi) and a cobalt-zebra (Maylandia callainos): Diagnostic imaging, clinical and histopathological study.

A female albino pindani (Chindongo socolofi) and female cobalt-zebra (Maylandia callainos) were referred to the Ornamental Fish Clinic with a soft tissue mass on their upper jaw. Also, during clinical examination, an unencapsulated intraoral soft tissue mass was observed in the cobalt-zebra cichlid. Digital radiography showed soft tissue masses and osteolytic lesions. Computed tomography (CT) imaging confirmed that osteolytic lesions occurred in skull bones of cobalt-zebra. Also, soft tissue mass invaded to the intraoral space. Histopathologically, a non-encapsulated, multilobulated and densely cellular neoplastic mass was identified. Extensive infiltration of neoplastic cells to the dermis and subcutaneous tissue was observed in both cases. The masses were composed of multiple coalescing nests, sheets, lobules and trabeculae of neoplastic cells within a fibrovascular stroma. The palisading basaloid neoplastic cells were markedly extended from the basal cells of the epidermis into the dermis and subcutis. The majority of the neoplastic cells had scant amounts of eosinophilic cytoplasm with indistinct margins, round to ovoid nuclei and typically a single small basophilic nucleolus. There was invasion of basaloid neoplastic cells into the bone tissue that caused osteolytic lesions. Mitotic figures were three to seven per high-power field. Epidermal basement membrane was periodic acid-Schiff (PAS) positive and neoplastic cell extension into the dermis was identified by the PAS staining. Fibrovascular stroma of the mass was also stained blue with Masson's trichrome. Based on histopathological and histochemical findings, the masses were diagnosed as infiltrative type of basal cell carcinoma (BCC).

Isolation of infectious laryngotracheitis virus in broiler chicken in Iran: First report.

In February 2019, a severe respiratory distress with co-infection of infectious laryngotracheitis (ILT) and Newcastle disease accompanied with Salmonella Enteritidis occurred in a broiler flock in the western region of Iran. Clinical signs included paralysis, torticollis, nasal discharge, conjunctivitis, gasping and respiratory rale with high mortality. At necropsy, caseous diphtheritic membrane adherent to the larynx and trachea was observed. Microscopically, syncytial cells formation with dense eosinophilic intranuclear inclusion bodies were main histopathological findings in tracheal tissues. Conventional polymerase chain reaction (PCR) for ICP4 gene amplification as a definitive diagnosis was utilized for the detection of ILT virus nucleic acid in suspected tracheal samples inoculated on to the chorioallantioc membrane of 11-day-old specific pathogen free (SPF) chicken eggs. Tracheal tissues taken from these SPF birds were positive by nested ILT PCR. In conclusion, because of no vaccination policy against ILT in broilers, the most probable scenario is that virus-laden dust or other fomites can be vectors and virus persistence and disease outbreak can be a sequel of wild virus introduction to the farm.

Occurrence of myxoma in a bearded dragon (Pogona vitticeps): Surgical and histopathological studies.

Myxoma is a benign mesenchymal tumor of skin and soft connective tissues considered to be of fibroblastic origin. A female bearded dragon (Pogona vitticeps) with a raised mass on the left side of the body was referred to the Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. The mass had become evident and grew more extensive within the previous 11-month period. The dragon was anesthetized with 5.00% isoflurane, and the mass was removed surgically. Tissue samples of the mass were fixed in 10.00% neutral buffered formalin, sectioned and stained with Hematoxylin and Eosin (H&E), Alcian blue (pH: 2.50), and periodic acid Schiff (PAS) staining techniques. The patient was administered meloxicam, lincomycin, and marbofloxacin. Histopathologically, the capsulated mass was composed of myxoid structure. Stellate-to-reticular cells distributed in a mucinous stroma containing few tiny blood vessels were seen. Myxoid cells displayed a PAS-positive cytoplasm. The myxomatous matrix was alcianophilic at the pH of 2.50. To our knowledge, this is the first report of surgery, treatment, and histopathological study of myxoma in a bearded dragon (P. vitticeps). The sutures were also removed 12 days after the operation. No new growth of the mass was observed three months following the surgical procedures.

Evaluation of multi-epitope recombinant protein as a candidate for a contraceptive vaccine.

Contraceptive vaccine (CV) is a valuable, non-invasive, and alternative method for purposeful contraception. Sperm antigens are useful targets for producing CVs due to their specialized expression in sperm. In this study, a recombinant protein containing three main sperm epitopes (IZUMO1, SACA3, and PH-20) was designed and evaluated as CV to control fertility in male mice. The chimeric recombinant protein was expressed and purified in E. coli. Male mice were immunized by 100 µg purified protein and sera were collected to assess IgG antibodies. Evaluating the reproductive performance, immunized male mice mated with normal-fertile female mice and mating rate and the number of newborns was studied. Immunized mice were sacrificed and necropsy and histopathology studies were conducted. The results revealed that the designed chimeric protein stimulated the immune system of the mice effectively. The level of IgG antibody was significantly higher in vaccinated mouse rather than control mouse. Eighty percent of the vaccinated mice became infertile and in the remaining ones, the number of children decreased to 4-6 offspring instead of 10-12 in normal mice. Histopathological studies showed that no organs including heart, brain, lung, liver, kidney and intestine were damaged. However, Normal spermatogenesis has been disrupted and necrotic spermatogonia cells were reported in Seminiferous tubules. We concluded that the designed chimeric protein containing IZUMO1, SACA3, and PH-20 epitopes can stimulate the immune system and cause male contraception without any side effects.